Shank 3 Mouse Model of Autism - Psychogenics...Shank 3 Mouse Model of Autism Confidential 2...
Transcript of Shank 3 Mouse Model of Autism - Psychogenics...Shank 3 Mouse Model of Autism Confidential 2...
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Shank 3 Mouse Model of Autism
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Background
• About 2% of people with autism carry harmful mutations in SHANK3, a protein that helps organize the connections between neurons
• Developed by Guoping Feng, Shank3 –Feng mice harbor a deletion of exons 13-16 of the PDZ domains leading to the deletion of the Shank3α and Shank3β isoforms and partial deletion of Shank3γ
• These mice exhibit some social, communicative, repetitive, and sensory processing abnormalities associated with autism spectrum disorder.
• Displays some social deficits• Shows a decrease in activity and locomotion compared to littermate controls,
including reduced rearing• Shows gait differences compared to littermate controls• Displays some anxiety-like behaviors• Shows reduced startle responses and increased prepulse inhibition of startle
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No Significant differences in Body Weight Between WT and KO Mice
Age (weeks)
5 6 7 8 9 10 11
Bo
dy W
eig
ht
(g)
10
15
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SHNAK3/F_WTSHNAK3/F_KO
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Behavioral Assessments
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KO Mice Show Deficits in Rotarod PerformanceL
ate
nc
y t
o F
all
(s
ec
)
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Shank3/F_WTShank3/F_KO
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Sp
eed
at
Fall
(rp
m)
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10
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Urine Open Field Test
• Male mice are placed in an Open Field where they are presented with the urine of an estrus female
• Activity, proximity to the urine, ultrasonic vocalizations, and urine marking by the male are recorded to determine social and function
• Urine spots developed with Nindyrin Spary
Estrous female urine spot
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KO Mice Show Decreased Activity in the Open Field Following Exposure to Female Urine
To
tal
Dis
tan
ce T
ravele
d (
cm
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SHNK3/F_WTSHNK3/F_KO
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Dis
tan
ce
Tra
ve
led
in
Ce
nte
r (c
m)
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SHNK3/F_WTSHNK3/F_KO
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Tim
e S
pen
t in
Ce
nte
r
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10
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Reciprocal Social Interaction
• Same-genotype, -sex, -age dyads are allowed to freely interact for 10 minutes
• Distance between mice, proximity, and interactions are measures of sociality
Front Back Side
Nu
mb
er
of
Inte
racti
on
s
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Tim
e S
pe
nt
Fo
llo
win
g (
sec)
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KO Mice Show Increased Grooming
Gro
om
ing
Bo
uts
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150
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KO Mice Show Decreased Startle Response
Me
an
Sta
rtle
Re
sp
on
se
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800
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1200
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KO Mice Show Increased Hindlimb Clasping
Mice are lifted gently by the tail with front limbs just above the surface
Clasping of hind legs is noted (normal is a spread in the hind legs)
Pe
rcen
t M
ice
Cla
sp
ing
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KO Mice Show Decreased Ultrasonic Vocalizations
Ult
ras
on
ic V
oca
liza
tio
ns
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Electrophysiology
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Impaired corticostriatal synaptic transmission in Shank3 KO
• Extracellular field potential responses (A) and whole-cell currents in medium spiny neurons (B) were evoked in dorsal striatum by
stimulating corpus callosum.
• Brain slices were prepared from male mice aged 10 weeks for extracellular field potential recordings (A) and 14 weeks for whole-
cell patch clamp recordings (B).
• Numbers in parenthesis show number of slices (cells); animals used.
A B
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Impaired NMDA-mediated synaptic transmission in dorsal striatum in Shank3 KO
• Whole-cell NMDA-mediated currents in medium spiny neurons were evoked in dorsal striatum by stimulating corpus
callosum in 14 weeks old male mice.
• Numbers in parenthesis show number of cells; animals used.
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Impaired synaptic transmission in nucleus accumbens in Shank3 KO
• Extracellular field potential responses were evoked in nucleus accumbens (NAc) by stimulating terminals within the
nucleus in 10 weeks old male mice.
NAcacp
Str
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Molecular markers
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Decreased expression of synaptic proteins in striatum and BNDF in cortex in Shank3 KO
• Expression of synaptic proteins in striatum and BDNF in cortex was assessed by qPCR in 16 weeks old male
Shank3 KO mice (n=10).